Fused bicyclic pyrimidine derivatives

ABSTRACT

The invention relates to compounds of the formula                    
     and to pharmaceutically acceptable salts thereof, wherein R 1 , R 2  and Z are as defined herein. The invention also relates to pharmaceutical compositions containing the compounds of formula I and to methods of using said compounds in the treatment of hyperproliferative diseases such as cancer.

This is a national stage filing under 35 USC Section 371 ofPCT/IB97/01393, filed Nov. 5, 1997, which claimed priority of U.S.Provisional Application Nos. 60/031,862 and 60/041,846 filed Nov. 27,1996 and Apr. 9, 1997, respectively.

BACKGROUND OF THE INVENTION

This invention relates to novel bicyclic pyrimidine derivatives that areuseful in the treatment of hyperproliferative diseases, such as cancers,in mammals. This invention also relates to a method of using suchcompounds in the treatment of hyperproliferative diseases in mammals,especially humans, and to pharmaceutical compositions containing suchcompounds.

Compounds that are useful in the treatment of hyperproliferativediseases are also disclosed in the following co-pending patentapplications: PCT international patent application numberPCT/IB97/100675 (filed Jun. 11, 1997), United States provisional patentapplication No. 60/028881 (filed Oct. 17, 1996), PCT internationalpatent application number PCT/IB97/00584 (filed May 22, 1997), U.S.patent application Ser. No. 08/653,786 (filed May 28, 1996), PCTinternational patent application publication number WO 96/40142(published Dec. 19, 1996), and PCT international patent applicationpublication number WO 95/23141 (published Aug. 31, 1995). Each of theforegoing United States and PCT international patent applications isincorporated herein by reference in its entirety.

It is known that a cell may become cancerous by virtue of thetransformation of a portion of its DNA into an oncogene (i.e., a genethat upon activation leads to the formation of malignant tumor cells).Many oncogenes encode proteins which are aberrant tyrosine kinasescapable of causing cell transformation. Alternatively, theoverexpression of a normal proto-oncogenic tyrosine kinase may alsoresult in proliferative disorders, sometimes resulting in a malignantphenotype.

Receptor tyrosine kinases are large enzymes that span the cell membraneand possess an extracellular binding domain for growth factors such asepidermal growth factor, a transmembrane domain, and an intracellularportion that functions as a kinase to phosphorylate specific tyrosineresidue in proteins and hence to influence cell proliferation. It isknown that such kinases are often aberrantly expressed in common humancancers such as breast cancer, gastrointestinal cancer such as colon,rectal or stomach cancer, leukemia, and ovarian, bronchial or pancreaticcancer. It has also been shown that epidermal growth factor receptor(EGFR), which possesses tyrosine kinase activity, is mutated oroverexpressed in many human cancers such as brain, lung, squamous cell,bladder, gastric, breast, head and neck, oesophageal, gynecological andthyroid cancers. Thus, it is believed that inhibitors of receptortyrosine kinases, such as the compounds of the present invention, areuseful as selective inhibitors of the growth of mammalian cancer cells.

It has also been shown that EGFR inhibitors may be useful in thetreatment of pancreatitis and kidney disease (such as proliferativeglomerulonephritis and diabetes-induced renal disease), and may reducesuccessful blastocyte implantation and therefore may be useful as acontraceptive. See PCT international application publication number WO95/19970 (published Jul. 27, 1995).

It is known that polypeptide growth factors such as vascular endothelialgrowth factor (VEGF) having a high affinity to the human kinaseinsert-domain-containing receptor (KDR) or the murine fetal liver kinase1 (FLK-1) receptor have been associated with the proliferation ofendothelial cells and more particularly vasculogenesis and angiogenesis.See PCT international application publication number WO 95/21613(published Aug. 17, 1995). Agents, such as the compounds of the presentinvention, that are capable of binding to or modulating the KDR/FLK-1receptor may be used to treat disorders related to vasculogenesis orangiogenesis such as diabetes, diabetic retinopathy, hemangioma, glioma,melanoma, Kaposi's sarcoma and ovarian, breast, lung, pancreatic,prostate, colon and epidermoid cancer.

SUMMARY OF THE INVENTION

The present invention relates to compounds of the formula

and to pharmaceutically acceptable salts thereof, wherein;

Z is a group of the formula

wherein n is an integer from 0 to 2 and p is an integer from 0 to 3;

R¹ is H, C₁-C₆ alkyl or —C(O)(C₁-C₆ alkyl);

R² is phenyl or 1H-indazol-5-yl, wherein said groups are optionallysubstituted by 1 to 3 R⁵ substituents, or R² is a group of the formula(Ii) or (Ij)

wherein p is an integer from 0 to 3 and n is an integer from 0 to 2;

or R¹ and R² are taken together to form a group of the formula (Ik)

wherein the dashed line indicates a single or double bond and m is aninteger from 0 to 4;

each R³ is independently H, —C(O)OR^(9,) or C₁-C₆ alkyl wherein saidalkyl is optionally substituted by halo, —OR⁹, —NR⁹R¹⁰ or —C(O)OR⁹;

R⁴ is R³ —OR⁹, or —NR⁹R¹⁰;

each R⁵ is independently halo, cyano, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆,alkynyl, —OR⁹, —NR⁹R¹⁰, nitro, or C₆-C₁₀ aryl wherein said alkyl,alkenyl, alkynyl and aryl R⁵ groups are optionally substituted by 1 to 3substituents independently selected from halo, nitro, C₁-C₄ alkyl and—OR⁹;

R⁶ and R⁷ are independently H or R⁵;

R⁸ is H, —SO₂(C₆-C₁₀ aryl), —(CH₂)_(q)(5-10 membered heterocyclyl),C₂-C₆ alkenyl, C₁-C₆ alkyl, —(CH₂)_(q)(CH₂)_(q)(C₁-C₆ alkoxy),—(CH₂)_(q)(C₁-C₆ alkoxy), —C(O)(C₁-C₆ alkoxy), or —SO₂(C₁-C₄ alkyl)wherein each q is independently an integer from 2 to 4;

each R⁹ and R¹⁰ is independently H or C₁-C₆ alkyl; and,

R¹¹ is trifluoromethyl, halo, nitro, —OR⁹, —NR⁹R¹⁰, cyano, C₁-C₄ alkyl,—S(O)_(x)R⁹ wherein x is an integer from 0 to 2,—C(O)OR⁹, —OC(O)(C₁-C₄alkyl), —C(O)NR⁹R¹⁰), —NR⁹C(O)(C₁-C₄ alkyl), —C(O)NHSO₂(C₁-C₄ alkyl),—NHCH₂C(O)NR⁹R¹⁰, —NHC(O)(C₁-C₄ alkoxy), —NHOC(O)(C₁-C₄ alkyl),—NR⁹OR¹⁰, anilino, pyrrolidinyl, piperidinyl, azido, guanidino,morpholino, phenyl, —C(O)(C₁-C₆ alkyl), benzenesulfonyl, allyl,thiophenyl, piperazinyl, 4-(C₁-C₄ alkyl)-piperazinyl, phenyftnio,benzenesulphonamido, 2-oxopyrrolidin-1-yl, 2,5dioxopyrrolidin-1-yl,phenoxy, benzoyloxy, benzoylamino, —(CH₂)_(w)O(CH₂)_(v)OR^(9,)—O(CH₂)_(w)O(CH₂)_(v)OR⁹, —O(CH₂)_(w)C(O)OR⁹, —O(CH₂)_(w)C(O)NR⁹R¹⁰,—(CH₂)_(w)S(CH₂)_(v)OR^(9,)—NH(CH₂)_(v)O(C₁-C₄ alkyl),—NH(CH₂)_(w)(C₆-C₁₀ aryl), —NHC(O)(CH₂)_(w)(C₁-C₄ alkoxy), or—O(CH₂)_(w)(C₆-C₁₀ aryl), wherein w is an integer from 1 to 4 and v isan integer from 2 to 4, and wherein the alkyl, heterocyclic, and arylmoieties of the foregoing R¹¹ groups are optionally substituted by 1 or2 substituents independently selected from the group consisting of halo,C₁-C₄ alkyl, —OR^(9,)—NR⁹R¹⁰, —C(O)OR⁹, —OC(O)(C₁—C₄ alkyl),—C(O)NR⁹R¹⁰, —NHC(O)(C₁-C₄ alkyl), nitro, imidazolyl, piperidino,morpholino, and piperazinyl;

with the proviso that where Z is a group of the formula (Ie), and R² isphenyl, then said phenyl is substituted by 1 to 3 substituentsindependently selected from C₁-C₆ alkyl, C₂-C₆ alkynyl and halo, and oneof R⁴ and R⁷ is halo or H and the other is as defined above; and,

with the further proviso that where Z is a group of the formula (Ia),(Ib), (Ic) or (Id), and R² is phenyl, then said phenyl is substituted byC₂-C₆ alkynyl.

Preferred compounds of formula I include those wherein R² is optionallysubstituted phenyl. More preferred are those compounds of formula Iwherein R² is phenyl substituted by C₂-C₆ alkynyl, in particularethynyl.

Other preferred compounds of formula I include those wherein Z is apyrrolo moiety of formula (Ie) or (If).

Other preferred compounds of formula I include those wherein R¹ and R²are taken together to form an indole or indoline moiety of formula (1k),

Specific preferred compounds of formula I include the following:

4-(6-Chloro-2,3dihydro-indol-1-yl)-7H-pyrrolo[2,3-d]pyrimidine;

4-(6-Methyl-2,3dihydro-indol-1-yl)-7H-pyrrolo[2,3-d]pyrimidine;

4 (6-Chloro-5-fluoro-2,3dihydryo-indol-1-yl)-7H-pyrrolo[2,3]pyrimidine:

1-(4-m-Tolylamino-pyrrolo[2,3-d]pyrimidin-7-yl)-ethanone;

4-(6Chloro-2,3-dihydro-indol-1-yl)-pyrido[3,4-d]pyrimidine;

4-(6-Bromo-5chloro-2,3dihydro-indol-1-yl)-pyrido[3,4-d]pyrimidine;

4-(6-Fluoro5chloro-2,3-dihydro-indol-1-yl)-pyrido[3,4-d]pyrimidine;

4(6lodo-2,3-dihydro-indol-1-yl)-pyrido[3,4-d]pyrimidine;

(7-Benzenesulfonyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-(3ethynyl-phenyl)-amine;

4-(6-Chloro-2,3-dihydro-indol-1-yl)-5H-pyrrolo[3,2-d]pyrimidin-6-ol;

(3-Ethynyl-phenyl)-[7(2morpholin-4-yl-ethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-amine;

(3-Ethynyl-phenyl)-[7-(2-methoxy-ethyl)-7H-pyrrolo[2,3-d]pyrmidin-4-yl]-amine;

(3Ethynyl-phenyl)-{7[2-(2-methoxy-ethoxy)ethyl]-7H-pyrrolo[2,3-d]pyrmidin4-yl}amine;

(7-Allyl-pyrrolo[2,3-d]pyrimidin-4-yl)-(3ethynyl-phenyl)-amine;

N-(5lodo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N-m-tolyl-acetamide;

4-(6-Chloro-2,3dihydro-indol-1-yl)6-methyl-pyrido[3,4-d]pyrimidine;

4-(6-Bromo-5-fluoro-2,3dihydro-indol-1-yl)6methyl-pyrido[3,4-d]pyrimidine;

4-(6-Chloro5-fluoro-2,3dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidine;

4-(6-lodo-2,3dihydro-indol-1-yl)-6-methyl-pyrido[3,4]pyrimidine;

4-(4-Bromo-7-methyl-2,3-dihydro-indol-1-yl)6-methyl-pyrido[3,4-d]pyrimidine;

4-(6-Bromo-7-methyl-2,dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidine;

4-(6,7-Dimethyl-2,3dihydro-indol-1-yl)pyrido[3,4-d]pyrimidine;

(3-Ethynyl-phenyl)pyrido[3,4-d]pyrimidin-4-yl-amine;

Benzo[b]thiophen-5yl-pyrido[3,4-d]pyrimidin4-yl-amine;

(3-Ethynyl-phenyl)-(5-tolyl-7H-pyrrolo[2,3-d]pyrimidin4-yl)-amine;

(3-Ethynyl-phenyl)-(5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine;

(3-Ethynyl-phenyl)-[5-(4-methoxy-phenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-amine;

(3-Ethynyl-phenyl)-[5-(3nitro-phenyl)7H-pyrrolo[2,3-d]pyrimidin-4-yl]-amine;

[5-(4-chloro-phenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-(3-ethynyl-phenyl)-amine;

(3-Bromo-phenyl)-(6-bromo-5-phenyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine;and the pharmaceutically acceptable salts of the foregoing compounds.

The invention also relates to a pharmaceutical composition for thetreatment of a hyperproliferative disorder in a mammal which comprises atherapeutically effective amount of a compound of formula I, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier. In one embodiment, said pharmaceutical compositionis for the treatment of cancer such as brain, lung, squamous cell,bladder, gastric, pancreatic, breast, head, neck, renal, kidney,ovarian, prostate, colorectal, oesophageal, gynecological or thyroidcancer. In another embodiment, said pharmaceutical composition is forthe treatment of a non-cancerous hyperproliferative disorder such asbenign hyperplasia of the skin (e.g., psoriasis) or prostate (e.g.,benign prostatic hypertropy (BPH)).

The invention also relates to a pharmaceutical composition for thetreatment of a hyperproliferative disorder in a mammal which comprises atherapeutically effective amount of a compound of formula I or apharmaceutically acceptable salt thereof, in combination with ananti-tumor agent selected from the group consisting of mitoticinhibitors, alkylating agents, anti-metabolites, intercalatingantibiotics, enzymes, topoisomerase inhibitors, biological responsemodifiers, anti-hormones, and anti-androgens, and a pharmaceuticallyacceptable carrier.

The invention also relates to a pharmaceutical composition for thetreatment of pancreatitis or kidney disease (including proliferativeglomerulonephritis and diabetes-induced renal disease) in a mammal whichcomprises a therapeutically effective amount of a compound of formula Ior a pharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier.

The invention also relates to a pharmaceutical composition for theprevention of blastocyte implantation in a mammal which comprises atherapeutically effective amount of a compound of formula I or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier.

The invention also relates to a pharmaceutical composition for treatinga disease related to vasculogenesis or angiogenesis in a mammal whichcomprises a therapeutically effective amount of a compound of formula Ior a pharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier. In one embodiment, said pharmaceutical compositionis for treating a disease selected from the group consisting ofdiabetes, diabetic retinopathy, hemangioma, glioma, melanoma, Kaposi'ssarcoma and ovarian, breast, lung, pancreatic, prostate, colon andepidermoid cancer.

The invention also relates to a method of treating a hyperproliferativedisorder in a mammal which comprises administering to said mammal atherapeutically effective amount of the compound of formula I or apharmaceutically acceptable salt thereof. In one embodiment, said methodrelates to the treatment of cancer such as brain, squamous cell,bladder, gastric, pancreatic, breast, head, neck, oesophageal, prostate,colorectal, lung, renal, kidney, ovarian, gynecological or thyroidcancer. In another embodiment, said method relates to the treatment of anon-cancerous hyperproliferative disorder such as benign hyperplasia ofthe skin (e.g., psoriasis) or prostate (e.g., benign prostatichypertropy (BPH)).

The invention also relates to a method for the treatment of ahyperproliferative disorder in a mammal which comprises administering tosaid mammal a therapeutically effective amount of a compound of formulaI, or a pharmaceutically acceptable salt thereof, in combination with ananti-tumor agent selected from the group consisting of mitoticinhibitors, alkylating agents, anti-metabolites, intercalatingantibiotics, enzymes, topoisomerase inhibitors, biological responsemodifiers, anti-hormones, and anti-androgens.

The invention also relates to a method of treating pancreatitis orkidney disease in a mammal which comprises administering to said mammala therapeutically effective amount of the compound of formula I or apharmaceutically acceptable salt thereof.

The invention also relates to a method of preventing blastocyteimplantation in a mammal which comprises administering to said mammal atherapeutically effective amount of the compound of formula I or apharmaceutically acceptable salt thereof.

The invention also relates to a method of treating diseases related tovasculogenesis or angiogenesis in a mammal which comprises administeringto said mammal an effective amount of a compound of formula I, or apharmaceutically acceptable salt thereof. In one embodiment, said methodis for treating a disease selected from the group consisting ofdiabetes, diabetic retinopathy, hemangioma, glioma, melanoma, Kaposi'ssarcoma and ovarian, breast, lung, pancreatic, prostate, colon andepidermoid cancer.

Patients that can be treated with compounds of the formula I, and thepharmaceutically acceptable salts of said compounds, according to themethods of this invention include, for example, patients that have beendiagnosed as having psoriasis, BPH, lung cancer, bone cancer, pancreaticcancer, skin cancer, cancer of the head and neck, cutaneous orintraocular melanoma, uterine cancer, ovarian cancer, rectal cancer,cancer of the anal region, stomach cancer, colon cancer, breast cancer,gynecologic tumors (e.g., uterine sarcomas, carcinoma of the fallopiantubes, carcinoma of the endometrium, carcinoma of the cervix, carcinomaof the vagina or carcinoma of the vulva), Hodgkin's disease, cancer ofthe esophagus, cancer of the small intestine, cancer of the endocrinesystem (e.g., cancer of the thyroid, parathyroid or adrenal glands),sarcomas of soft tissues, cancer of the urethra, cancer of the penis,prostate cancer, chronic or acute leukemia, solid tumors of childhood,lymphocytic lymphonas, cancer of the bladder, cancer of the kidney orureter (e.g., renal cell carcinoma, carcinoma of the renal pelvis), orneoplasms of the central nervous system (e.g., primary CNS lymphona,spinal axis tumors, brain stem gliomas or pituitary adenomas).

The term “halo”, as used herein, unless otherwise indicated, meansfluoro, chloro, bromo or iodo. Preferred halo groups are fluoro, chloroand bromo.

The term “alkyl”, as used herein, unless otherwise indicated, includessaturated monovalent hydrocarbon radicals having straight, cyclic orbranched moieties. It is understood that for cyclic moieties at leastthree carbon atoms are required in said alkyl group.

The term “alkoxy”, as used herein, unless otherwise indicated, includesO-alkyl groups wherein “alkyl”, is as defined above.

The term “aryl”, as used herein, unless otherwise indicated, includes anorganic radical derived from an aromatic hydrocarbon by removal of onehydrogen, such as phenyl or naphthyl.

The term “5-10 membered heterocyclyl”, as used herein, unless otherwiseindicated, includes aromatic and non-aromatic heterocyclic groupscontaining one or more heteroatoms each selected from O, S and N,wherein each heterocyclic group has from 5-10 atoms in its ring system.The heterocyclic groups include benzo-fused ring systems and ringsystems substituted with one or more oxo moieties. An example of a 5membered heterocyclic group is thiazolyl, and an example of a 10membered heterocyclic group is quinolinyl. Examples of non-aromaticheterocyclic groups are pyrrolidinyl, piperidino, morpholino,thiomorpholino and piperazinyl. Examples of aromatic heterocyclic groupsare pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl,tetrazolyl, furyl, thienyl, isoxazolyl and thiazolyl. Heterocyclicgroups having a fused benzene ring include benzimidazolyl.

The phrase “pharmaceutically acceptable salt(s)”, as used herein, unlessotherwise indicated, includes salts of acidic or basic groups which maybe present in the compounds of formula I. The compounds of formula Ithat are basic in nature are capable of forming a wide variety of saltswith various inorganic and organic acids. The acids that may be used toprepare pharmaceutically acceptable acid addition salts of such basiccompounds of formula I are those that form non-toxic acid additionsalts, i.e., salts containing pharmacologically acceptable anions, suchas the hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate,bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate,salicylate, citrate, acid citrate, tartrate, pantothenate, bitartrate,ascorbate, succinate, maleate, gentisinate, fumarate, gluconate,glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate,ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate [i.e.,1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts.

Those compounds of the formula I that are acidic in nature, are capableof forming base salts with various pharmacologically acceptable cations.Examples of such salts include the alkali metal or alkaline earth metalsalts and particularly, the sodium and potassium salts.

Certain compounds of formula I may have asymmetric centers and thereforeexist in different enantiomeric forms. This invention relates to the useof all optical isomers and stereoisomers of the compounds of formula Iand mixtures thereof. The compounds of formula I may also exist astautomers. This invention relates to the use of all such tautomers andmixtures thereof.

DETAILED DESCRIPTION OF THE INVENTION

The preparation of the compounds of the present invention is illustratedin the following Schemes 1 and 2.

The compounds of the present invention are readily prepared according tosynthetic methods familiar to those skilled in the art. Such methods aredisclosed in PCT international patent application publication number WO96/40142 (published Dec. 19, 1996), referred to above, and in PCTinternational application publication numbers WO 95/19774 (publishedJul. 27, 1995) and WO 95/19970 (published Jul. 27, 1995), each of whichis incorporated herein by reference.

Scheme 1 illustrates the coupling of the bicyclic compound of formula IIwith the amine of formula III to provide the compound of formula I. Inthe compounds of formulas II and III, X is hydroxy or chloro and Z. R¹and R² are as defined above. In general, the compound of formula II iscoupled with the amine of formula III in a solvent, such as a C₁-C₆alcohol, dimethylformamide (DMF), N-methylpyrrolidin-2-one (NMP),chloroform, acetonitrile, tetrahydrofuran (THF), dimethylsulfoxide(DMSO), 1,4-dioxane or pyridine, optionally in the presence of a base,such as pyridine or triethylamine, and optionally in the presence ofpyridine hydrochloride as a catalyst, under an inert atmosphere, such asdry nitrogen gas, at a temperature of from ambient to refluxtemperature, preferably 80-125° C., for a period of about 2 hours to 72hours.

The bicyclic compound of formula II is prepared according to syntheticmethods known to those skilled in the art. Such methods are disclosed inWO 95/19774 and WO 95/19970, referred to above. Other methods ofpreparing the compounds of formula II are also disclosed in PCTinternational application publication number WO 92/12718 (published Aug.6, 1992), A. Petric et al., Fur Chemie 114, 615-624 (1983) and NucleicAcids Research, v. 12, no. 2 (1984). Where the compound of formula IIIis an optionally substituted indole or indoline moiety, such compoundscan be prepared according to one or more methods known to those skilledin the art. Such methods are described in PCT international patentapplication publication number WO 95/23141, referred to above, and in W.C. Sumpter and F. M. Miller, “Heterocylic Compounds with Indole andCarbazole Systems,” in volume 8 of “The Chemistry of HeterocyclicCompounds”, Interscience Publishers Inc., New York (1954). Optionalsubstituents can be included as appropriate before or after the couplingstep illustrated in Scheme 1. Prior to the coupling step, primary andsecondary amino moieties (other than the amine of formula III) arepreferably protected using a nitrogen protecting group known to thoseskilled in the art. Such protecting groups and their use are describedin T. W. Greene and P. G. M. Wuts, “Protective Groups in OrganicSynthesis,” Second Edition, John Wiley & Sons, New York, 1991.

Scheme 2 illustrates the cyclization of the compound of formula IV(wherein Me means methyl) to provide the compound of formula V. Thisillustrates the introduction of the Z moiety in the compound of formulaI after the amino (—NR¹R²) moiety has been coupled with the pyrimidinering. .While Scheme 2 illustrates the formation of apyrrolo[3,2-d]pyrimidine ring system, other ring systems can be made byone or more analogous methods disclosed in WO 90/19774, referred toabove. In Scheme 2, the compound of formula IV, in an alcoholic solvent,such as ethanol, is catalytically hydrogenated using an appropriatecatalyst, such as 10% palladium on carbon, under an H₂ atmosphere atambient temperature for a period of about three hours to provide thecompound of formula V. Optional substituents can be introduced into thecompound of formula V, as appropriate, according to methods known tothose skilled in the art, to provide the desired compound of formula I.

The compounds of the present invention may have asymmetric carbon atoms.Such diasteromeric mixtures can be separated into their individualdiastereomers on the basis of their physical chemical differences bymethods known to those skilled in the art, for example, bychromatography or fractional crystallization. Enantiomers can beseparated by converting the enantiomeric mixtures into a diastereomricmixture by reaction with an appropriate optically active compound (e.g.,alcohol), separating the diastereomers and converting (e.g.,hydrolyzing) the individual diastereomers to the corresponding pureenantiomers. All such isomers, including diastereomer mixtures and pureenantiomers are considered as part of the invention.

The compounds of formula I that are basic in nature are capable offorming a wide variety of different salts with various inorganic andorganic acids. Although such salts must be pharmaceutically acceptablefor administration to animals, it is often desirable in practice toinitially isolate the compound of formula I from the reaction mixture asa pharmaceutically unacceptable salt and then simply convert the latterback to the free base compound by treatment with an alkaline reagent andsubsequently convert the latter free base to a pharmaceuticallyacceptable acid addition salt. The acid addition salts of the basecompounds of this invention are readily prepared by treating the basecompound with a substantially equivalent amount of the chosen mineral ororganic acid in an aqueous solvent medium or in a suitable organicsolvent, such as methanol or ethanol. Upon careful evaporation of thesolvent, the desired solid salt is readily obtained. The desired acidsalt can also be precipitated from a solution of the free base in anorganic solvent by adding to the solution an appropriate mineral ororganic acid.

Those compounds of the formula I that are acidic in nature, are capableof forming base salts with various pharmacologically acceptable cations.Examples of such salts include the alkali metal or alkaline-earth metalsalts and particularly, the sodium and potassium salts. These salts areall prepared by conventional techniques. The chemical bases which areused as reagents to prepare the pharmaceutically acceptable base saltsof this invention are those which form non-toxic base salts with theacidic compounds of formula I. Such non-toxic base salts include thosederived from such pharmacologically acceptable cations as sodium,potassium calcium and magnesium, etc. These salts can easily be preparedby treating the corresponding acidic compounds with an aqueous solutioncontaining the desired pharmacologically acceptable cations, and thenevaporating the resulting solution to dryness, preferably under reducedpressure. Alternatively, they may also be prepared by mixing loweralkanolic solutions of the acidic compounds and the desired alkali metalalkoxide together, and then evaporating the resulting solution todryness in the same manner as before. In either case, stoichiometricquantities of reagents are preferably employed in order to ensurecompleteness of reaction and maximum yields of the desired finalproduct.

The active compounds of this invention are potent inhibitors of the erbBfamily of oncogenic and protooncogenic protein tyrosine kinases such asepidermal growth factor receptor (EGFR), erbB2, HER3, or HER4 and thusare all adapted to therapeutic use as antiproliferative agents (e.g.,anticancer) in mammals, particularly in humans. In particular, thecompounds of this invention are useful in the prevention and treatmentof a variety of human hyperproliferative disorders such as malignant andbenign tumors of the liver, kidney, bladder, breast, gastric, ovarian,colorectal, prostate, pancreatic, lung, vulval, thyroid, hepaticcarcinomas, sarcomas, glioblastomas, head and neck, and otherhyperplastic conditions such as benign hyperplasia of the skin (e.g.,psoriasis) and benign hyperplasia of the prostate (e.g., BPH). It is, inaddition, expected that a compound of the present invention may possessactivity against a range of leukemias and lymphoid malignancies.

The active compounds may also be useful in the treatment of additionaldisorders in which aberrant expression ligand/receptor interactions oractivation or signalling events related to various protein tyrosinekinases, are involved. Such disorders may include those of neuronal,glial, astrocytal, hypothalamic, and other glandular, macrophagal,epithelial, stromal, and blastocoelic nature in which aberrant function,expression, activation or signalling of the erbB tyrosine kinases areinvolved. In addition, compounds of formula I may have therapeuticutility in inflammatory, angiogenic and immunologic disorders involvingboth identified and as yet unidentified tyrosine kinases that areinhibited by compounds of the formula I.

The in vitro activity of the active compounds in inhibiting the receptortyrosine kinase (and thus subsequent proliferative response, e.g.,cancer) may be determined by the following procedure.

Activity of the active compounds, in vitro, can be determined by theamount of inhibition of the phosphorylation of an exogenous substrate(e.g., Lys₃-Gastrin or polyGluTyr (4:1) random copolymer (I. Posner etal., J. Biol. Chem. 267 (29), 20638-47 (1992)) on tyrosine by epidermalgrowth factor receptor kinase by a test compound relative to a control.Affinity purified, soluble human EGF receptor (96 ng) is obtainedaccording to the procedure in G. N. Gill, W. Weber, Methods inEnzymology 146, 82-88 (1987) from A431 cells (American Type CultureCollection, Rockville, Md.) and preincubated in a microfuge tube withEGF (2,μg/ml) in phosphorylation buffer +vanadate (PBV: 50 mM HEPES, pH7.4; 125 mM NaCl; 24 mM MgCl₂; 100 μM sodium orthovanadate), in a totalvolume of 10 μl, for 20-30 minutes at room temperature. The testcompound, dissolved in dimethylsulfoxide (DMSO), is diluted in PBV, and10 μl, is mixed with the EGF receptor IEGF mix, and incubated for 10-30minutes at 30° C. The phosphorylation reaction is initiated by additionof 20 μl³³P-ATP/substrate mix (120 μM Lys₃-Gastrin (sequence in singleletter code for amino acids, KKKGPWLEEEEEAYGWLDF), 50 mM Hepes pH 7.4,40 μM ATP, 2 μCi γ-[³³P]-ATP) to the EGFr/EGF mix and incubated for 20minutes at room temperature. The reaction is stopped by addition of 10μl stop solution (0.5 M EDTA, pH 8; 2 mM ATP) and 6 μl 2N HCl. The tubesare centrifuged at 14,000 RPM, 4° C., for 10 minutes. 35 μl ofsupernatant from each tube is pipetted onto a 2.5 cm circle of WhatmanP81 paper, bulk washed four times in 5% acetic acid, 1 liter per wash,and then air dried. This results in the binding of substrate to thepaper with loss of free ATP on washing. The [³³P] incorporated ismeasured by liquid scintillation counting. Incorporation in the absenceof substrate (e.g., lys₃-gastrin) is subtracted from all values as abackground and percent inhibition is calculated relative to controlswithout test compound present.

Such assays, carried out with a range of doses of test compounds, allowthe determination of an approximate IC₅₀ value for the in vitroinhibition of EGFR kinase activity. The compounds of the formula I thatwere tested using the procedure described above exhibited IC₅₀ values inthe range of 0.0001-30 μM.

Activity of the active compounds, in vivo, can be determined by theamount of inhibition of tumor growth by a test compound relative to acontrol. The tumor growth inhibitory effects of various compounds aremeasured according to the methods of Corbett T. H., et al. “TumorInduction Relationships in Development of Transplantable Cancers of theColon in Mice for Chemotherapy Assays, with a Note on CarcinogenStructure”, Cancer Res., 35, 2434-2439 (1975) and Corbett, T. H., etal., “A Mouse Colon-tumor Model for Experimental Therapy”, CancerChemother. Rep. (Part 2)”, 5, 169-186 (1975), with slight modifications.Tumors are induced in the left flank by s.c. injection of 1×10⁶ logphase cultured tumor cells (human MDA-MB-468 breast or human HN5 headand neck carcinoma cells) suspended in 0.10 ml RPMI 1640. Aftersufficient time has elapsed for the tumors to become palpable (2-3 mm indiameter) the test animals (athymic mice) are treated with activecompound (formulated by dissolution in DMSO typically at a concentrationof 50 to 100 mg/mL followed by 1:9 dilution into saline or,alternatively, 1:9 dilution into 0.1% Pluronic® P105 in 0.9% saline) bythe intraperitoneal (ip) or oral (po) routes of administration twicedaily i.e., every 12 hours) for 5 consecutive days. In order todetermine an anti-tumor effect, the tumor is measured in millimeterswith Vernier calipers across two diameters and the tumor size (mg) iscalculated using the formula: Tumor weight=(length×[width]²)/2,according to the methods of Geran, R. I., et al. “Protocols forScreening Chemical Agents and Natural Products Against Animal Tumors andOther Biological Systems”, Third Edition, Cancer Chemother. Rep., 3,1-104 (1972). Results are expressed as percent inhibition, according tothe formula: Inhibition(%)=(TuW_(control)−TuW_(test))/TuW_(control×)100%. The flank site oftumor implantation provides reproducible dose/response effects for avariety of chemotherapeutic agents, and the method of measurement (tumordiameter) is a reliable method for assessing tumor growth rates. Thetitle compounds of the experimental examples of this case that arecompounds of the formula I all exhibited, when tested in the aboveassay, percent inhibition values greater than 50% at 10 μM.

Other methods of assessing the activity of the compounds of the presentinvention are referred to in PCT international application publicationnumber WO 95/21613 (published Aug. 17, 1995) which incorporated hereinby reference.

Administration of the active compounds can be effected by any methodthat enables delivery of the compounds to the site of action (e.g.,cancer cells). These methods include oral routes, intraduodenal routes,parenteral injection (including intravenous, subcutaneous,intramuscular, intravascular or infusion), topical administration, etc.

The amount of the active compound administered will be dependent on thesubject being treated, the severity of the disorder or condition, therate of administration and the judgement of the prescribing physician.However, an effective dosage is in the range of about 0.001 to about 100mg per kg body weight per day, preferably about 1 to about 35mg/kg/day,in single or divided doses. For a 70 kg human, this would amount toabout 0.05 to about 7 g/day, preferably about 0.2,to about 2.5 g/day. Insome instances, dosage levels below the lower limit of the aforesaidrange may be more than adequate, while in other cases still larger dosesmay be employed without causing any harmful side effect, provided thatsuch larger doses are first divided into several small doses foradministration throughout the day.

The active compound may be applied as a sole therapy or may involve oneor more other anti-tumour substances, for example those selected from,for example, mitotic inhibitors, for, example vinblastine; alkylatingagents, for example cis-platin, carboplatin and cyclophosphamide;anti-metabolites, for example 5-fluorouracil, cytosine arabinoside andhydroxyurea, or, for example, one of the preferred anti-metabolitesdisclosed in European Patent Application No. 239362 such asN-(5[N-(3,4-dihydro-2-methyl-4-oxoquinazolin6-ylmethyl)-N-methylamino]2-thenoyl)-L-glutamicacid; intercalating antibiotics, for example adriamycin and bleomycin;enzymes, for example interferon; and anti-hormones, for exampleanti-estrogens such as Nolvadex™ (tamoxifen) or, for exampleanti-androgens such as Casodex™(4-cyano-3-(4-fluomphenylsulphonyl)-2hydroxy-2methyl-3′-(trifluoromethyl)propionanilide).Such conjoint treatment may be achieved by way of the simultaneous,sequential or separate dosing of the individual components of thetreatment.

The pharmaceutical composition may, for example, be in a form suitablefor oral administration as a tablet, capsule, pill, powder, sustainedrelease formulations, solution, suspension, for parenteral injection asa sterile solution, suspension or emulsion, for topical administrationas an ointment or cream or for rectal administration as a suppository.The pharmaceutical composition may be in unit dosage forms suitable forsingle administration of precise dosages. The pharmaceutical compositionwill include a conventional pharmaceutical carrier or excipient and acompound according to the invention as an active ingredient. Inaddition, it may include other medicinal or pharmaceutical agents,carriers, adjuvants, etc.

Exemplary parenteral administration forms include solutions orsuspensions of active compounds in sterile aqueous solutions, forexample, aqueous propylene glycol or dextrose solutions. Such dosageforms can be suitably buffered, if desired.

Suitable pharmaceutical carriers include inert diluents or fillers,water and various organic solvents. The pharmaceutical compositions may,if desired, contain additional ingredients such as flavorings, binders,excipients and the like. Thus for oral administration, tabletscontaining various excipients, such as citric acid may be employedtogether with various disintegrants such as starch, alginic acid andcertain complex silicates and with binding agents such as sucrose,gelatin and acacia. Additionally, lubricating agents such as magnesiumstearate, sodium lauryl sulfate and talc are often useful for tabletingpurposes. Solid compositions of a similar type may also be employed insoft and hard filled gelatin capsules. Preferred materials, therefor,include lactose or milk sugar and high molecular weight polyethyleneglycols. When aqueous suspensions, or elixirs are desired for oraladministration the active compound therein may be combined with varioussweetening or flavoring agents, coloring matters or dyes and, ifdesired, emulsifying agents or suspending agents, together with diluentssuch as water, ethanol, propylene glycol, glycerin, or combinationsthereof.

Methods of preparing various pharmaceutical compositions with a specificamount of active compound are known, or will be apparent, to thoseskilled in this art. For examples, see Remington's PharmaceuticalSciences, Mack Publishing Company, Easter, Pa., 15th Edition (1975).

The following examples illustrate the preparation of the compounds ofthe invention. In the following examples, “Me” means methyl and “Et”means ethyl.

EXAMPLE 1

4-(6-Chloro2,3dihydro-indol-1-yl)-7H-pyrrolo[2,3-d]pyrimidine

To 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (10.0 g, 0.065 mol) in drypyridine (90 ml) was added 6chloro2,3dihydroindol-1-yl amine (14.8 g,0.078 mol), and the mixture was heated in an 85° C. oil bath for 2 days.The reaction was cooled to ambient temperature and concentrated invacuo. The resulting residue was purified by flash chromatography onsilica gel (375 g, 40 mm mesh) using 5% MeOH/CH₂Cl₂ to afford the titlecompound as a pink-orange solid (1.1 g, 4.3%): HRMS: Calculated271.0750, Found 271.0729; anal. RP18-HPLC RT: 4.88 minutes.

The above compound was dissolved in minimal methanol and a solution ofHCl in (HCl(g) bubbled into 2 ml Et₂O) was added dropwise until themixture remained cloudy. The precipitated HCl salt was dried in vacuo,washed once with Et₂O, and dried in vacuo to constant mass (MP: 266° C.(dec)).

EXAMPLES 2-3

The compounds of Examples 2 and 3 were made according to the method ofExample 1 from 4-chloro-7H-pyrrolo[2,3-d]pyrimidine and the appropriateamine starting material.

Example RP18- LC/MS no. R % Yield HPLC RT (M⁺) 2 6-Methyl 23 4.62 251 36-Chloro-5-fluoro 23 4.66 289

EXAMPLE 4

1-(4-m-Tolylamino-pyrrolo[2,3-d]pyrimidin-7: yl)-ethanone

Following the procedure described in Example 1,(7H-Pyrrolo[2,3-d]pyrimidin4-yl)-m-tolyl-amine was prepared from4-chloro-7H-pyrrolo[2,3-d]pyrimidine and m-toluidine (34%): HRMS:Calculated 225.1140, Found 225.1131; anal. RP18-HPLC RT: 3.45 min; HClsalt MP: 219° C.

To (3methyl-phenyl)-(7H-pyrrolo[2,3-d]pyrimidin4-yl)-amine (0.168 g,0.75 mmol) dissolved in hot acetonitrile (7 ml) was added sodium hydride(36 mg, 0.90 mmol, 60% dispersion in mineral oil). After stirring atambient temperature for 0.75 hour, acetyl chloride (0.11 ml, 1.5 mmol)was added and stirring continued for 48 hours. The mixture wasconcentrated in vacuo, triturated in hot ethyl acetate, and filtered.The filtrate was concentrated in vacuo to give an orange solid residue.The solid was triturated in CH₂Cl₂ and filtered to afford the titlecompound as a light yellow solid (0.11 g, 55%): LC-MS: 267 (MH⁺); anal.RP18-HPLC RT: 3.53 minutes.

EXAMPLE 5

4-(6-Chloro-2,3dihydro-indol-1-yl)-pyrido[3,4-d]pyrimidine

To a suspension of 4-hydroxy-pyrido[3,4-d]pyrimidine (0.103 g, 0.70mmol) in dry pyridine (2 ml) cooled in an ice-water bath was addeddropwise trifluoroacetic anhydride (0.20 ml, 1.4 mmol). After stirringfor 0.5 hour, 6-chloroindoline (0.10 g, 0.66 mmol) and pyridine (0.14 g,1.81 mmol) in dry DMF (1.5 ml) was added dropwise. The cold bath wasallowed to warm to ambient temperature and then heated at 70° C. for 3hours. The reaction was cooled to ambient temperature and then added tomethylene chloride (150 mL). The organic layer was washed with saturatedsodium carbonate and water, and then dried over sodium sulfate. Thesolvent was removed by rotary evaporation and the residue purified bycolumn chromatography (silica gel, 9/2/1-CH₂Cl₂/hexanes/methanol) togive a pale yellow residue (0.048 g, 28%): MP: 194-6° C.; LC/MS: 283(MH⁺).

EXAMPLES 6-8

The compounds of examples 6-8 were made according to the method ofExample 5 from 4-chloro-pyrido[3,4-d]pyrimidine and appropriate aminestarting materials.

Example # R Yield HPLC RT LC/MS (M⁺) 6 6-Bromo-5-fluoro 45 3.64 359 76-Chloro-5-fluoro 315 8 6-Iodo 389

EXAMPLE 9

(7-Benzenesulfonyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-(3-ethynyl-phenyl)-amine

To 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (1.0 g, 0.0065 mol) in dry THF(10 ml) under nitrogen at −78° C. was added dropwise via syringe over 15minutes n-butyllithium (2.5 M in hexane; 2.88 ml, 0.0072 mol). Thecooling bath was removed and the solution was stirred for 1 hour. Theresulting pyrrolo anion precipitated as a very fine white solid in acloudy colorless solution. After the suspension was re-cooled to −78°C., benzenesulfonyl chloride (1.26 g, 0.0072 mol) was added neat viasyringe. The resulting yellow mixture was allowed to warm slowly toambient temperature overnight. The grey-white suspension was poured into2% aqueous sodium bicarbonate (50 mL) and extracted with diethyl ether(4×20 mL). The combined extracts were washed with water and dried(potassium carbonate) and evaporated to give a light amber oil whichcrystallized from diethyl ether. The product was collected by filtrationto 1.4 g (74%) of white solid: LC-MS=294 (MH⁺); anal. RP18-HPLC RT: 4.40minutes

The above compound was dissolved in MeOH and m-aminophenyl acetylene(0.159 g, 0.0013 mol), and the mixture was heated in an 85° C. oil bathfor 2 days. The reaction was cooled to ambient temperature andconcentrated in vacuo. The residue was triturated with diethyl ether toproduce a white solid (0.234 g. 92%): LC-MS: 375 (MH⁺); anal. RP18-HPLCRT: 3.48 minutes.

EXAMPLE 10

4-(6Chloro-2,3dihydro-indol-1-yl)-5H-pyrrolo[3,2-d]pyrimidin-6-ol

To a solution of4(6Chloro-2,3-dihydro-indol-1-yl)-5-amino-6-methylacetyl-pyrimidine (541mg, 1.55 mmol) in 40 mL of ethanol was added 25 mol % of 10% palladiumon carbon (125 mg) and 0.11 mL of 1N HCl (1.55 mmol). The mixture washydrogenated for 3 hours at 3.4 atm (50 psi)(H₂). The reaction isfiltered through Celite® and concentrated in vacuo. The brown residuewas slurried in methanol and the white solid was filtered off (279 mg,63%): LC-MS: 287 (M⁺); anal. RP18-HPLC RT: 5.61 minutes; MP: 250° C.(dec).

EXAMPLE 11

(3-Ethynyl-phenyl)-[7-(2-morpholin-4-yl-ethyl)-7H-pyrrolo[2,3-d]pyrimidin4-yl]-amine

To a solution of 184 mg (1.4 mmol) of 4-(2-hydroxyethyl)morpholine in 10mL of toluene was added 276 mg (2.0 mmol) of anhydrous potassiumcarbonate and then 32 mg (1.3 mmol) of 97% sodium hydride. After 30minutes, 343 mg (1.0 mmol) of sulfonylated4-chloro-7H-pyrrolo[2,3-d]pyrimidine was added and the reaction washeated at 100° C. for 2 hours. The reaction was then partitioned betweenethyl acetate and water and the aqueous layer was extracted with twoadditional portions of ethyl acetate. The combined organic phase waswashed with water, dried over magnesium sulfate and concentrated invacuo. The residue was chromatographed on silica gel using 10%methanol/methylene chloride to give an amber oil (140 mg, 55%): LC-MS267 (M⁺).

The above compound was dissolved in MeOH and m-aminophenyl acetylene(0.123 g, 0.001 mol), and the mixture was heated in a sealed tube in a120° C. oil bath for 12 hours. The reaction was cooled to ambienttemperature and concentrated in vacuo. The residue was triturated withdiethyl ether to produce a white solid (0.135 g, 74%): LC-MS=348 (MH⁺);anal. RP18-HPLC RT: 3.33 minutes.

EXAMPLE 12

(3-Ethynyl-phenyl)-[7-(2-methoxy-ethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-amine

Following a procedure analogous to that described in Example 11, thetitle product was prepared in 81% yield from4-chloro7-(2-methoxy-ethyl)-7H-pyrrolo[2,3-d]pyrimidine (1.0 eq) andm-aminophenyl acetylene (1.2 eq) in methanol: MP: 240-241° C.; LC-MS:292 (MH⁺); anal. RP18-HPLC RT: 4.16 minutes.

EXAMPLE 13

(3-Ethynyl-phenyl)-{7-[2-(2-methoxy-ethoxy)-ethyl]-7H-pyrrolo[2,3-d]pyrimidin4-yl}amine

Following a procedure analogous to that described in Example 11, thetitle product was prepared in 81% yield from4-chloro-7-[2-(2-methoxy-ethoxy)-ethyl]-7H-pyrrolo[2,3-d]pyrimidine (1.0eq) and m-aminophenyl acetylene (1.2 eq) in methanol: MP: 240-241° C.;LC-MS: 336 (M⁺); anal. RPP18-HPLC RT: 4.29 minutes.

EXAMPLE 14

(7-Allyl-pyrrolo[2,3-d]pyrimidin-4-yl)-(3-ethynyl-phenyl)-amine

To 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (1.3 g: 8.5 mmol) in dry THF (30ml) was added sodium hydride (1.0 g, 0.25 mmol, 60% dispersion inmineral oil). After stirring at ambient temperature for 1 hour, allyliodide (0.93 ml, 10 mmol) was added and stirring continued for 48 hours.The mixture was concentrated in vacuo, triturated in hot ethyl acetate,and filtered. The filtrate was concentrated in vacuo to give an orangesolid residue. The solid was triturated in CH₂Cl₂ and filtered to afford4-chloro-7-allyl-pyrrolo[2,3-d]pyrimidine as a light yellow powder (0.58g, 36%): TS-MS: 194 (MH⁺).

To 4-chloro-7-allyl-pyrrolo[2,3-d]pyrimidine (0.5 g, 2.6 mmol) in dryMeOH (5 ml) was added m-aminophenyl acetylene (0.36 g, 3.1 mmol). Thesuspension was heated in a sealed pressure tube at 125° C. for 20 hours.The reaction was cooled to ambient temperature and concentrated invacuo. The resulting oil was purified by flash chromatography on silicagel (50 g, 40 mm mesh) using 3% MeOH/CH₂Cl₂ to afford the title compoundas a yellow powder (0.29 g, 41%): TS-MS: 275 (MH⁺); anal. RP18-HPLC RT:4.62 minutes.

EXAMPLE 15

N-(5-lodo-7H-pyrrolo[2,3-d]pyrimidin4-yl)-N-m-tolyl-acetamide

To (3-methyl-phenyl)-(7H-pyrrolo[2,3-d]pyrimidin4-yl)-amine (0.75 g, 3.4mmol) dissolved in hot acetonitrile (30 ml) was added sodium hydride(0.16 g, 4.0 mmol, 60% dispersion in mineral oil). After stirring atambient temperature for 0.75 hour, acetyl chloride (0.48 ml, 6.7 mmol)was added and the mixture was stirred for 48 hours. The mixture wasconcentrated in vacuo, triturated in hot ethyl acetate, and filtered.The filtrate was concentrated in vacuo to give an orange solid residue.The solid was purified by flash chromatography on silica gel (13 g, 40mm mesh) using 1:3 ethyl acetate/hexanes to afford the title product asa yellow solid (0.21 g): TS-MS: 309 (MH⁺).

To 1-(4-m-tolylamino-pyrrolo[2,3-d]pyrimidin-7-yl)-ethanone (0.21 g,0.79 mmol) in dry CH₂Cl₂ (5 ml) and dry MeOH (2 ml) was added Na₂CO₃(0.17 g, 1.6 mmol). After stirring at ambient temperature for 0.75 hour,N-iodosuccinimide (0.35 g, 1.6 mmol) was added. The mixture was stirredat ambient temperature for 48 hours then concentrated in vacuo. Theresidue was diluted with CH₂Cl₂ and H₂O. The H₂O phase was extractedonce with CH₂Cl₂. The organic phase was washed twice with H₂O, driedover Na₂SO₄, and concentrated in vacuo. The resulting residue waspurified by flash chromatography on silica gel (11 g, 40 mm mesh) using2% MeOH/CH₂Cl₂ to afford the title compound as a yellow solid (30 mg):TS-MS: 393 (MH⁺); anal. RP18-HPLC RT: 3.42 minutes.

EXAMPLE 16

4-(6Chloro-2,3-dihydro-indol-1-yl)-6methyl-pyrido[3,4-d]pyrimidine

6Methyl-pyrido[3,4-d]pyrimid-4one (200 mg, 1.24 mmol), polymer-supportedtriphenylphosphine (2.06 g of 3.0 mmol Pig resin, 6.20 mmol) andanhydrous carbon tetrachloride (1.20 mL, 12.40 mmol) were combined in1,2-dichloroethane (6 mL). The mixture was heated to 60° C. under anatmosphere of dry N₂(g) for 18 hours. 6-Chloroindoline (1.1 eq.) wasadded and heating was continued at 60° C. for another 18 hours. Thetriphenyphosphine-bearing resin was filtered off and washed severaltimes with chloroform. The filtrate and washings were concentrated invacuo and preparative reversed-phase (C18) chromatography utilizing agradient of 15% to 70% MeCN/pH4.5, 50 mM NH₄OAc followed bylyophilization of the appropriate fractions afforded the title product(30%) as its free-base: MP: 232-234° C.; LC-MS: 297 (MH⁺); anal.RP-HPLC: 4.33 minutes.

EXAMPLES 17-19

The compounds of examples 17-19 were made according to the method ofExample 16 from 4-chloro6-methyl-pyrido[3,4-d]pyrimidine and theappropriate amine starting materials.

Example RP18-HPLC no. R Yield RT LC/MS (M⁺) 17 6-Bromo-5-fluoro 45 3.64359 18 6-Chloro-5-fluoro 315 19 6-Iodo 389

EXAMPLE 20

4-(4-Bromo-7-methyl-2,3-dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidinehydrochloride salt

The title product was prepared from 6methyl-pyrido[3,4-d]pyrimid-4-one(1.0 eq.) and 4-bromo-7-methyl-indoline (1.5 eq.) according to theprocedure described for Example 16. The crude product from the filtratewas flash chromatographed on silica using EtOAc/hexanes/MeOH (9:2:1) toafford the free base which was converted to the hydrochloride salt asdescribed for Example 23 (33% yield): MP: 232-244° C.; LC-MS: 355, 357(MH⁺); anal. RP-HPLC: 5.20 minutes.

EXAMPLE 21

4-(6-Bromo-7-methyl-2,3-dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidinehydrochloride salt

The title product was prepared from 6-methyl-pyrido[3,4-d]pyrimid-4-one(1.0 eq.) and 6-bromo-7-methyl-indoline (1.5 eq.) according to theprocedure described for Example 16. The crude product from the filtratewas flash chromatographed on silica using EtOAc/hexanes/MeOH (9:2:1) toafford the free base which was converted to the hydrochloride salt asdescribed for Example 23 (34% yield): MP 212-229° C.; LC-MS: 355, 357(MH⁺); anal. RP-HPLC: 4.90 minutes.

EXAMPLE 22

4-(6-Bromo-5-fluoro-2,3dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidine hydrochloride salt

The title product was prepared from 6-methyl-pyrido[3,4-d]pyrimid-4-one(1.0 eq.) and 6-bromo-5-fluoro-indoline (1.5 eq.) according to theprocedure described for Example 16. The crude product from the filtratewas flash chromatographed on silica using 2% MeOH/98% CH₂Cl₂ to affordthe free base which was converted to the hydrochloride salt as describedfor Example 23 (36% yield): MP: 262-264° C.; LC-MS: 359, 361 (MH⁺);anal. RP-HPLC: 4.83 minutes.

EXAMPLE 23

4-(6,7-Dimethyl-2,3-dihydro-indol-1-yl)pyrido[3,4-d]pyrimidine

To 4-chloro-pyrido[3,4-d]pyrimidine (200 mg, 1.21 mmol) in isopropanol(3 mL) was added 6,7-dimethylindoline (211 mg, 1.44 mmol) and pyridine(190 mg, 2.41 mmol). The mixture was heated to reflux under anatmosphere of dry N₂(g) for 6hours. Solvent was removed in vacuo and theresidue was dissolved in CHCl₃ and washed with saturated aqueous Na₂CO₃.The organic phase was dried over Na₂SO₄, concentrated in vacuo, andflash chromatographed on silica in 45% acetone/hexanes to afford 60 mgof 4-(6,7dimethyl-2,3-dihydro-indol-1-yl)-pyrido[3,4-d]pyrimidine(LC-MS: 278 (MH⁺). This material was dissolved in a minimum volume of10% MeOH in CH₂Cl₂, and 1 mole equivalent of HCl in diethyl ether wasadded dropwise with stirring. The mixture was diluted with diethyl ether(4 volumes) and the precipitated HCl salt of the product was filteredand dried in vacuo (58 mg): MP: 248° C.; GEMS: 277 (M⁺); anal. RP-HPLC:4.06 minutes.

EXAMPLE 24

(3-Ethynyl-phenyl)-pyrido[3,4-d]pyrimidin-4-yl-amine

To 4-chloro-pyrido[3,4-d]pyrimidine (250 mg, 1.50 mmol) inN-methylpyrrolidin-2-one (0.5 mL) was added 3ethynylaniline (212 mg,1.81 mmol) and pyridine (237 mg, 3.0 mmol). The mixture was heated to80° C. under an atmosphere of dry N₂(g) for 3 hours. The reactionmixture was dissolved in CHCl₃ and washed with saturated aqueous Na₂CO₃,and brine. The organic phase was dried over Na₂SO₄, concentrated invacuo, and flash chromatographed on silica with a gradient of 40% to 70%acetone/hexanes to afford 120 mg of product. This material wasprecipitated as its hydrochloride salt by dissolution in minimal CHCl₃,titration with HCl (1 eq.) in diethyl ether, and dilution with diethylether. The yellow salt was filtered and dried in vacuo (133 mg): MP233-235° C.; LC-MS: 247 (MH⁺); anal. RP-HPLC: 3.45 minutes.

EXAMPLE 25

Benzo[b]thiophen-5-yl-pyrido[3,4-d]pyrimidin-4-yl-amine

To 4-chloro-pyrido[3,4-d]pyrimidine (250 mg, 1.50 mmol) inN-methylpyrrolidin-2-one (0.5 mL) was added benzo[b]thiophen-5-yl-amine(270 mg, 1.81 mmol) and pyridine (237 mg, 3.0 mmol). The mixture washeated to 80° C. under an atmosphere of dry N₂(g) for 3 hours. Thereaction mixture was dissolved in CHCl₃ and washed with saturatedaqueous Na₂CO₃, and brine. The organic phase was dried over Na₂SO₄(s),concentrated in vacuo, and flash chromatographed on silica in 40% to 70%acetone/hexanes to afford 180 mg of product. This material wasprecipitated as its hydrochloride salt by dissolution in minimal CHCl₃,tritration with HCl (1 eq.) in diethyl ether, and dilution with diethylether. The yellow salt was filtered and dried in vacuo (188 mg): MP:280-282° C.; LC-MS: 279 (MH⁺); anal. RP-HPLC: 3.63 minutes.

EXAMPLE 26

(3-Ethynyl-phenyl)-(5-p-tolyl-7H-pyrrolo[2,3-d]pyrimidin4-yl)-amine

To a solution of7-Benzenesulfonyl-4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (400 mg,0.953 mmol) in 8 mL of toluene was added this (benzylidene acetone)dipalladium chloroform adduct (40 mg, 10% by weight), 4-methyl benzeneboronic acid (260 mg, 1.91 mmol), sodium carbonate solution (2.86 ml of1 M solution) and 3.0 ml of ethanol (absolute). The mixture was refluxedfor seven hours. The reaction mixture was filtered through Celite®, andthe Celite® was washed with ethyl acetate. The filtrate was poured intoa separatory funnel was washed with saturated sodium bicarbonate (100mL, 2 times). The organic layer was dried over magnesium sulfate andconcentrated in vacuo. The resulting green black oil was stirred inmethanol (5 mL) and KOH (107 mg, 1.91 mmol in 3 mL of water) was added.After three hours, the reaction mixture was concentrated and the residuedissolved in water. The solution was acidified with 1 N HCl andextracted with ethyl acetate (50 ml, 3 times). The organic layer wasdried over MgSO₄ and concentrated in vacuo. The resulting oil waschromatographed on silica gel (180 g, 40 μm) using 30% ethylacetate/hexanes to provide the product as a white solid (225 mg 92%):TS-MS: 244, 246 (MH⁺). To 4-chloro-(5-p-tolyl)-7H-pyrrolo[2,3-d]pyrimidine (200 mg, 0.821 mmol) in dry methanol (4 mL) was addedm-aminophynel acetylene (106 mg, 0.903 mmol). The suspension was heatedin a sealed tube at 120° C. for 18 hours. The reaction was cooled toambient temperature and concentrated in vacuo to a brown solid. Theresidue was chromatographed over silica gel (140 g, 40 μm) using 40%ethyl acetate/hexanes to provide the title compound as a light yellowsolid (82 mg, 32%): MP: 245-247° C.; anal. RP18-HPLC RT: 6.120 minutes.

EXAMPLE 27

(3-Ethynyl-phenyl)-(5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrimidin4-yl]-amine

Following the procedure described in Example 26, the title compound wasprepared in 40% yield from7-Benzenesulfonyl-4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine andthiophene-2-boronic acid followed by coupling with m-aminophenylacetylene (27 mg, 0.233 mmoles) in methanol: MP: 230-232° C.; TS-MS: 317(MH⁺); anal. RP18-HPLC RT: 5.614 minutes.

EXAMPLE 28

(3-Ethynyl-phenyl-[5-(4-methoxy-phenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-amine

Following the procedure described in Example 26,4-chloro-[5-(4-methoxy-phenyl)]-7H-pyrrolo[2,3-d]pyrimidine, theintermediate, was made in 74% yield from7-benzenesulfonyl-4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine and4-methoxy benzene boronic acid: TS-MS: 260 (MH⁺). The title compound wassynthesized by coupling the intermediate with m-aminophenyl acetylene in75%yield: MP: 207-210° C.; FAB MS: 341 (MH⁺); anal. RP18-HPLC RT=5.640minutes.

EXAMPLE 29

(3-Ethynyl-phenyl)-[5-3-nitro-phenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-amine

Following the procedure described in Example 26,4-chloro-[5(4-methoxy-phenyl)]-7H-pyrrolo[2,3-d]pyrimidine, theintermediate, was made in 74% yield from7-benzenesulfonyl-4-chloro-5iodo-7H-pyrrolo[2,3-d]pyrimidine and 3nitrobenzene boronic acid: TS-MS: 260 (MH⁺). The title compound wassynthesized from coupling the intermediate with m-aminophenyl acetylenein 15% yield: MP 189-190° C.; TSMS: 469 (MH⁺); anal. RP18-HPLC RT=4.67minutes.

EXAMPLE 30

[5-(4-chloro-phenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-(3-ethynyl-phenyl)-amine

Following the procedure described in Example 26,4-chloro-[5-(4-methoxy-phenyl)]-7H-pyrrolo[2,3-d] pyrimidine, theintermediate, was made in 74% yield from7-benzenesulfonyl-4-chloro-5iodo-7H-pyrrolo[2,3-d]pyrimidine and4-chloro benzene boronic acid: TS-MS: 260 (MH⁺). The title compound wassynthesized by coupling the intermediate with m-aminophenyl acetylene in33% yield: MP: 225-227° C.; TSMS: 398 (MH⁺); anal. RP18-HPLC RT=6.45minutes.

EXAMPLE 31

(3-Bromo-phenyl)-(6bromo-5-phenyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine

Utilizing an analogous procedure as described in Example 26,4-chloro-(5-phenyl)-7H-pyrrolo[2,3-d] pyrimidine, the intermediate, wasmade in 74% yield from7-benzenesulfonyl-4-chloro-5iodo-7H-pyrrolo[2,3-d]pyrimidine and 3-bromobenzene boronic acid: TS-MS: 230 (MH⁺) This second intermediate wascoupled with 3-bromoaniline in methanol in a sealed tube to provide(3-bromo-phenyl)-[5-(4-chloro-phenyl)7H-pyrrolo[2,3-d]pyrimidin-4-yl]-aminein 75% yield: TSMS: 365 (MH+).(3-bromo-phenyl)-[5-(4-chloro-phenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-aminewas brominated with N-bromosuccinimide impregnated onto silica gel. Thereaction was complete after stirring at room temperature under nitrogenfor 1.5 hours. The reaction was filtered and concentrated in vacuo toprovide the title compound as a gray solid in 28% yield (128 mg): MP.:dec 300° C.; TSMS: 445/447 (MH⁺/MH⁺2); anal. RP1 8-HPLC RT=7.223minutes.

What is claimed is:
 1. A compound of the formula

or a pharmaceutically acceptable salt thereof, wherein: Z is a group ofthe formula

wherein n is an integer from 0 to 2 and p is an integer from 0 to 3; R¹is H, C₁-C₆ alkyl or —C(O)(C₁ C₆ alkyl); R² is 1H-indazol-5-yl, whereinsaid groups are optionally substituted by 1 to 3 R⁵ substituents, or R²is a group of the formula (Ii) or (Ij)

wherein p is an integer from 0 to 3 and n is an integer from 0 to 2; orR¹ and R² are taken together to form a group of the formula (Ik)

wherein the dashed line indicates a single or double bond and m is aninteger from 0 to 4; each R³ is independently H, —C(O)OR⁹, or C₁-C₆alkyl wherein said alkyl is optionally substituted by halo, —OR⁹,—NR⁹R¹⁰, or —C(O)OR⁹; R⁴ is R³, —OR⁹, or —NR⁹R¹⁰; each R⁵ isindependently halo, cyano, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₅ alkynyl,—OR⁹, —NR⁹R¹⁰, nitro, or C₆-C₁₀ aryl wherein said alkyl, alkenyl,alkynyl and aryl R⁵ groups are optionally substituted by 1 to 3substituents independently selected from halo, nitro, C₁-C₄ alkyl and—OR⁹; R⁶ and R⁷ are independently H or R⁵; R⁸ is H, —SO₂(C₆-C₁₀aryl),(CH₂)_(q)(5-10 membered heterocyclyl), C₂-C₆ alkenyl, C₁-C₆ alkyl,—(CH₂)_(q)O(CH₂)_(q)(C₁-C₆ alkoxy), —(CH₂)_(q)(C_(1-C) ₆ alkoxy),—C(O)(C₁-C₆ alkoxy), or —SO₂(C₁-C₄ alkyl) wherein each q isindependently an integer from 2 to 4; each R⁹ and R¹⁰ is independently Hor C₁-C₆ alkyl; and, R¹¹ is trifluoromethyl, halo, nitro, —OR⁹, —NR⁹R¹⁰,cyano, C₁-C₄ alkyl, —S(O)_(x)R⁹ wherein x is an integer from 0 to2,—C(O)OR⁹, —OC(O)(C₁-C₄ alkyl), —C(O)NR⁹R¹⁰, —NR⁹C(O)(C₁-C₄ alkyl),—C(O)NHSO₂(C₁-C₄ alkyl), —NHCH₂C(O)NR⁹R¹⁰, —NHC(O)(C₁-C₄ alkoxy),—NHOC(O)(C₁-C₄ alkyl), —NR⁹OR¹⁰, anilino, pyrrolidinyl, piperidinyl,azido, guanidino, phenyl, —C(O)(C₁-C₆ alkyl), benzenesulfonyl, allyl,thiophenyl, morpholino, piperazinyl, 4-(C₁-C₄ alkyl)piperazinyl,phenylthio, benzenesulphonamido, 2-oxopyrrolidin-1-yl,2,S-dioxopyrrolidin-1-yl, phenoxy, benzoyloxy, benzoylamino,—CH₂)_(w)O(CH₂)_(v)OR⁹, —O(CH₂)_(w)O(CH₂)_(v)OR⁹, —O(CH₂)_(w)C(O)OR⁹,—O(CH₂)_(w) C(O)NR⁹R¹⁰, —(CH₂)_(w)S(CH₂)_(v)OR⁹, —NH (CH₂)_(v)O(C₁-C₄alkyl), —NH(CH₂)_(w)(C₆-C₁₀ aryl), —NHC(O)(CH₂)_(w)(C₁-C₄ alkoxy), or—O(CH₂)_(w)(C₆-C₁₀, aryl), wherein w is an integer from 1 to 4 and v isan integer from 2 to 4, and wherein the alkyl, heterocyclic, and arylmoieties of the foregoing R¹¹ groups are optionally substituted by 1 or2 substituents independently selected from the group consisting of halo,C₁-C₄ alkyl, —OR⁹, —NR⁹R¹⁰, —C(O)OR⁹, —OC(O)(C₁-C₄ alkyl), —C(O)NR⁹R¹⁰,—NHC(O)(C₁-C₄ alkyl), nitro, imidazolyl, piperidino, morpholino, andpiperazinyl.
 2. A compound selected from the group consisting of:4-(6-Chloro-2,3dihydro-indol-1-yl)-7H-pyrrolo[2,3-d]pyrimidine;4-(6-Methyl-2,3-dihydro-indol-1-yl)-7H-pyrrolo[2,3-d]pyrimidine;4-(6-Chloro-5-fluoro-2,3dihydro-indol-1-yl)-7H-pyrrolo[2,3-d]pyrimidine;4-(6-Chloro-2,3dihydro-indol-1-yl)-pyrido[3,4-d]pyrimidine;4-(6-Bromo-5-chloro-2,3-dihydro-indol-1-yl)-pyrido[3,4-d]pyrimidine;4-(6-Fluoro-5-chloro-2,3dihydro-indol-1-yl)-pyrido[3,4-d]pyrimidine;4-(6-lodo-2,3-dihydro-indol-1-yl)pyrido[3,4-d]pyrimidine;4-(6-Chloro-2,3dihydro-indol-1-yl)-5H-pyrrolo[3,2-d]pyrimidin-6-ol;4-(6-Chloro-2,3′-dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidine;4-(6-Bromo-5-fluoro-2,3-dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidine;4-(6-Chloro-5-fluoro-2,3-dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidine;4-(6-lodo-2,3dihydro-indol-1-yl)6-methyl-pyrido[3,4-d]pyrimidine;4-(4-Bromo-7-methyl-2,3dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidine;4-(6-Bromo-7-methyl-2,3-dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidine;4-(6,7-Dimethyl-2,3dihydro-indol-1-yl)pyrido[3,4-d]pyrimidine;Benzo[b]thiophen-5-yl-pyrido[3,4-d]pyrimidin-4-yl-amine; and thepharmaceutically acceptable salts of the foregoing compounds.
 3. Apharmaceutical composition for the treatment of a hyperproliferativedisorder in a mammal which comprises a therapeutically effective amountof a compound of claim 1 and a pharmaceutically acceptable carrier. 4.The pharmaceutical composition of claim 3 wherein saidhyperproliferative disorder is cancer.
 5. The pharmaceutical compositionof claim 4 wherein said cancer is brain, lung, kidney, renal, ovarian,squamous cell, bladder, gastric, pancreatic, breast, head, neck,oesophageal, gynecological, prostate, colorectal or thyroid cancer. 6.The pharmaceutical composition of claim 3 wherein saidhyperproliferative disorder is noncancerous.
 7. The pharmaceuticalcomposition of claim 6 wherein said disorder is a benign hyperplasia ofthe skin or prostate.
 8. A pharmaceutical composition for the treatmentof a hyperproliferative disorder in a mammal which comprises atherapeutically effective amount of a compound of claim 1 in combinationwith an anti-tumor agent selected from the group consisting of mitoticinhibitors, alkylating agents, anti-metabolites, intercalatingantibiotics, enzymes, topoisomerase inhibitors, biological responsemodifiers, anti-hormones, and anti-androgens, and a pharmaceuticallyacceptable carrier.
 9. A pharmaceutical composition for the treatment ofpancreatitis or kidney disease in a mammal which comprises atherapeutically effective amount of a compound of claim 1 and apharmaceutically acceptable carrier.
 10. A pharmaceutical compositionfor the blastocyte implantation in a mammal which comprises atherapeutically effective amount of a compound of claim 1 and apharmaceutically acceptable carrier.
 11. A pharmaceutical compositionfor treating a disease related to vasculogenesis or angiogenesis in amammal which comprises a therapeutically effective amount of a compoundof claim 1 and a pharmaceutically acceptable carrier.
 12. Thepharmaceutical composition of claim 11 wherein said disease is selectedfrom the group consisting of diabetes, diabetic retinopathy, hemangioma,glioma, melanoma, Kaposi's sarcoma, ovarian cancer, breast cancer, lungcancer, pancreatic cancer, prostate cancer, colon cancer and epidermoidcancer.
 13. A method of treating a hyperproliferative disorder in amammal which comprises administering to said mammal a therapeuticallyeffective amount of a compound of claim
 1. 14. The method of claim 12wherein said hyperproliferative disorder is cancer.
 15. The method ofclaim 14 wherein said cancer is brain, lung, squamous cell, renal,kidney, ovarian, bladder, gastric, pancreatic, breast, head, neck,oesophageal, prostate, colorectal, gynecological or thyroid cancer. 16.The method of claim 13 wherein said hyperproliferative disorder isnoncancerous.
 17. The method of claim 16 wherein said disorder is abenign hyperplasia of the skin or prostate.
 18. A method for thetreatment of a hyperproliferative disorder in a mammal which comprisesadministering to said mammal a therapeutically effective amount of acompound of claim 1 in combination with an anti-tumor agent selectedfrom the group consisting of mitotic inhibitors, alkylating agents,anti-metabolites, intercalating antibiotics, enzymes, topoisomeraseinhibitors, biological response modifiers, anti-hormones, andanti-androgens.
 19. A method of treating pancreatitis or kidney diseasein a mammal which Comprises administering to said mammal atherapeutically effective amount of a compound of claim
 1. 20. A methodof preventing blastocyte implantation in a mammal which comprisesadministering to said mammal a therapeutically effective amount of acompound of claim
 1. 21. A method for treating a disease related tovasculogenesis or angiogenesis in a mammal which comprises administeringto said mammal a therapeutically effective amount of a compound ofclaim
 1. 22. The method of claim 21 wherein said disease is selectedfrom the group consisting of diabetes, diabetic retinopathy, hemangioma,glioma, melanoma, Kaposi's sarcoma, ovarian cancer, breast cancer, lungcancer, pancreatic cancer, prostate cancer, colon cancer and epidermoidcancer.